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From: Abizar Lakdawalla[SMTP:abizarl@innogenex.com]
Sent: Wednesday, April 28, 1999 12:44 PM
To: mogdie@innogenex.com
Subject: BrdU Staining
Try pretreating the slides with 0.1N HCl for 10-30 min, before you start
with the immunostaining procedure. At least with the IIB5 anti-BrdU clone
the staining intensity is significantly enhanced.
Matt
www.innogenex.com
Histonetters:
I have been using BrdU labeling of cells for quite
some time in my studies. Lately, I've been having
problems with the staining quality of the tissues from
some animals. I use BrdU from Sigma and anti-BrdU
from Becton Dickinson. Tissues collected are fixed
with Zinc Formalin (Anatech) for a minimum of 48h,
processed together, and stained in large batches using
capillary gap immuno. But about 10-15% of the animals
don't stain for BrdU at all, not even in the positive
control tissues (hair follicles, squamous epithelium).
There just seems to be no pattern to this! It's not
associated with any type of study exposure. Does
anyone have any suggestions as to what else can
contribute to this? I'm wondering if trying antigen
retrieval may help, but if the antigen isn't even
there, what next!?
===
Catherine "Katie" Bresee Bennett
Laboratory for Experimental Pathology
Department of Veterinary Pathology
Michigan State University